Objective To explore the expressions of adiponectin and adiponectin receptor 2 (AdipoR2) in the liver of patients with hepatitis B virus (HBV)-related liver failure and their clinical implications in the pathogenesis of liver failure.Methods The healthy controls (HC),patients with chronic hepatitis B (CHB) and patients with HBV-related liver failure (HBV-LF) were enrolled in the study.Each group had 20 participants.The liver tissues from 11 CHB patients who were diagnosed by liver biopsy,6 patients with HBV-LF and 6 liver donors during liver transplantation were collected.Histological specimens were observed by hematoxylin-eosin staining and Masson trichrome staining under microscope.The mRNA and protein expressions of adiponectin and AdipoR2 in the liver were measured by semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR) and immunohistochemical staining,respectively.The level of serum adiponectin was detected by enzymelinked immunsorbent assay.Serum biochemical parameters and HBV DNA levels were also measured.The pairwise comparison between groups was done by Student-Newman-Keuls and mann whitney U test.The relationship between two variables was analyzed using Spearman correlation.Results The level of serum adiponectin in HBV-LF group [(0.86 ± 0.15) ng/mL] was higher than those in HC [(0.59±0.15) ng/mL] and CHB groups [(0.62±0.13) ng/mL,Z=3.963,Z=3.774,both P＜0.01)],but showing no difference between CHB and HC groups (P＞0.05).The expressions of adiponectin and AdipoR2 mRNA in the liver were significantly higher in HBV-LF group (0.251 ±0.028 and 0.223 ± 0.021,respectively) than those in HC (0.091 ± 0.018 and 0.072 ± 0.020,respectively) and CHB (0.121±0.019 and 0.097±0.017,respectively) groups (q=18.428,17.069,19.807,18.673,respectively; all P＜0.01).Also,the expressions of adiponectin and AdipoR2 mRNA in CHB group were significantly higher than those in HC group (q=3.895,3.860,both P＜0.05).The expressions of adiponectin and AdipoR2 proteins in the liver were significantly higher in HBV-LF group (8.482±0.772 and 7.654±0.272,respectively) than those in HC (4.045± 0.815 and 2.804±0.623,respectively) and CHB (5.545±0.613 and 4.775±0.458,respectively) groups (q=15.327,11.542; Z=2.802,3.266; respectively; all P＜0.01).Also,the expressions of adiponectin,AdipoR2 proteins in CHB group were significantly higher than those in HC group (q=5.894,Z=3.266,both P＜0.01).In HBV-LF group,serum adiponectin levels as well as the expressions of adiponectin mRNA and protein in the liver were negatively correlated with serum albumin (r=-0.815,-0.886,-0.943; P＜0.01 or P＜0.05),but positively correlated with serum alanine aminotransferase (r=0.701,0.886,0.943; P＜0.01 or P＜0.05).The expression of AdipoR2 mRNA in the liver was negatively correlated with serum albumin (r=-0.943,P＜0.01),but positively correlated with serum aspartate aminotransferase (r=0.829,P＜0.05).Conclusions The expressions of adiponectin and AdipoR2 are up-regulated during HBV infection,especially in patients with HBV-LF,which might reflect the degree of necroinflammation in the liver.

OBJECTIVE To investigates the effects of imperatorin on the oxidative stress in the cerebral cortex and hippocampus after focal cerebral ischemia/reperfusion injury. METHODS Transient focal cerebral ischemia/reperfusion model in male Sprague-Dawley rats was induced by 2 h middle cerebral artery occlusion followed by 24 h reperfusion. Imperatorin (1.25 and 2.5 mg·kg- 1) or vehicle were administered intraperitoneally at 1, 5 and 9 h after the onset of ischemia. At 24 h after reperfusion, the biomarkers of oxidative stress such as the levels of reactive oxygen species (ROS), lipid peroxidation products malondialdehyde (MDA), nitric oxide (NO) and total antioxidant capacity (T-AOC), the activities of inducible nitric oxide synthase (iNOS), superoxide dismutase (SOD) and catalase (CAT) in the cerebral cortex and hippocampus were observed. We also assessed the nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and the NAD(P)H-quinone oxidoreductase 1 (NQO-1) protein expression by Western blot. RESULTS As compared to vehicle-treated animals, imperatorin treatment significantly reduced the ROS, MDA, NO levels and iNOS activity, increased T-AOC and the activities of SOD and CAT. Furthermore, imperatorin treatment also significantly induced the nuclear translocation of Nrf2, enhanced the protein expression of HO-1 and NQO-1 in the cerebral cortex and hippocampus. CONCLUSION Our findings indicate that imperatorin can protect the brain against the excessive oxidative stress induced by cerebral ischemia/reperfusion through activation of Nrf2 signaling pathway.

ObjectiveTo investigate the effects and molecular mechanism of simvastatin in liver fibrosis model of non-alcoholic fatty liver disease (NAFLD) in vivo and hepatic stellate cell in vitro.MethodsFirstly,the rat liver fibrosis model of NAFLD was established by high-fat diet administration and intervened with simvastatin.The expression of endothelial nitric oxide synthase (eNOS),inducible nitric oxide synthase (iNOS) and Collagen Ⅰ at mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Secondly,quiescent phenotype of LX-2 cell line was induced by promoting adipocyte differentiation medium in vitro,and then the quiescent phenotype of LX-2 cell were treated with transforming growth factor β1(TGF-β1),Nitroso-L-arginine methyl ester (L-NAME) which was NOS inhibitor,simvastatin,TGFβ1 with simvastatin,and L-NAME with simvastatin separately.The changes of eNOS,iNOS,αsmooth muscle actin (α-SMA) and Collagen Ⅰexpressions at mRNA and protein level were determined by RT-PCR and Western blot.ResultsAs modeling time extended,the expressions of eNOS in rat's liver tissue of model group at mRNA and protein level decreased gradually,however the expression of iNOS and Collagen Ⅰ at mRNA and protein level increased gradually,compared with normal control group and the differences were statistically significant (P ＜0.05 and 0.01).By 24weeks,the expressions of eNOS in rat's liver tissue of simvastatin group at mRNA and protein level were increased,the expression of iNOS at mRNA and protein level were decreased and the expression of Collagen Ⅰ at mRNA and protein level were decreased (all P ＜0.05).The expression of eNOS in rat's liver tissue of model group negatively correlated with the expression of Collagen Ⅰ at mRNA and protein level (all P ＜0.01).The expression of iNOS positively correlated with that of Collagen Ⅰ at mRNA and protein level (all P ＜0.01).In LX-2 cell culture,L-NAME inhibited the activation of LX-2,reduced eNOS and iNOS expression and increased α-SMA and CollagenⅠexpression,consistent with the role of TGF-β1.Simvastatin could directly increase the eNOS expression both in quiescent and activated LX-2 cells,decrease iNOS expression,maintain quiescent phenotype and inhibit its activation.ConclusionsSimvastatin ameliorated the genesis and progression of liver fibrosis by increasing eNOS expression in LX-2 cells and reducing iNOS,α-SMA and Collagen Ⅰ expression.

OBJECTIVETo introduce the framework of evidence-based practice with a case of castration-resistant prostate cancer (CRPC) as an example.

METHODSA clinical question was formulated according the clinical scenario. A systematic search was conducted for the published literature in the databases of PubMed, EMBASE, Cochrane Library, Clinical Trial Registries, and Web of Knowledge up to Dec 2014. The identified literature was reviewed for quality appraisal before the evidence was applied to clinical practice.

RESULTSThe treatment was effective and the patient achieved disease remission.

CONCLUSIONEvidence-based practice should be integrated with clinical scenario, current evidence, and patients' willingness, and follow a systematic framework.

For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Objective To explore the biological activities and the MR imaging signal intensities (SIs) characteristics of differentiations of neural-like cells induced by superparamagnetic iron oxide (SPIO)
and green fluorescent protein (GFP) double-labeled bone marrow-derived mesenchymal stem cells (BMSCs) in vitro. Methods GFP-BMSCs were labeled with different concentrations of SPIO in vitro (the concentration of the A, B, C and D group was 25, 50, 75 and 100 ug/ml, respectively;the E group without labels of SPIO served as the control group). The Prussian blue stainings were used to detect the labeling rates of SPIO. The iron contents of cells were measured by atomic absorption spectrometer. The CCK8 experiments were used to detect the cell proliferation rates. The cell cycles were detect by PCR. Each of the A-E groups had a test tube with 1 × 108 cells. All test tubes underwent T2* weighted imaging (T2*WI) and susceptibility weighted imaging (SWI) in a MR imaging scanner. The optimal group was defined by comparing the measurements of SIs between T2*WI and SWI. The optimal group and the E group together induced the differentiations of osteogenesis and neural-like cells. The stainings of alizarin red, osteocalcin and Nissl, NeuN, and NF-200 were performed at 72 hours. Real-time quantitative PCR was used to detect the expression levels of RNA in tub3, nestin, NSE, MAP-2 and Syt1. The positive staining rates and the expression levels of RNA were compared between the two groups. Finally, SWI was used to analyse the changes of SIs. One-way analysis of variance (ANOVA) was used to the multi-group comparison. Using least significant difference (LSD) test to analyse the comparisons between the multi-groups. Results The labeling rates of the A-D groups were 100%. The iron contents of cells in the A-E groups were (14.36 ± 7.61), (21.73±3.42), (30.54±8.73), (33.65±9.62), and (2.31±0.32) pg/cell, respectively. The iron contents of cells in the A-D groups were significantly higher than those in the E group ( F=3.852, P=0.003). There was no significant difference between the C and D groups (P=0.267). In all groups, the D group had the lowest OD value in the CCK-8 experiments (3.18 ± 0.46). In the A-E groups, the changes of SIs in SWI were significantly lower than those in T*2 WI. There was no significant difference in SIs of SWI between the C group (145.89±14.31) and the D group (127.37±12.21). Except the comparison between the group C and D, the comparisons between all the groups had significant differences (P<0.001). The percentages of SI attenuations in SWI and T*2 WI were 48.15% and 69.34%, respectively. The proportions of non-neurons cells and the positive rates of Nissl's stainings in group C and E had no significant differences (P>0.05). The expression levels of tub3, nestin and NSE were significantly higher before than after induced differentiations (P<0.01). SIs of SWI had no significant difference between before and after induced differentiations in the C group (t=1.26, P=0.236). Conclusions SPIO and GFP double-labeled BMSCs can induce neural-like cells without influencing biologic activities. MR SIs are decreased by the increase of SPIO concentrations in cells. SWI was the most sensitive sequence. The SIs of SWI has no differnce between before and after induced differentiations.

To study the preventive effect of Grifola frondosa on nonalcoholic steatohepatitis (NASH). The rat model of NASH was established by feeding high-fat diets for 12 weeks and intervened with 0.5 g · kg(-1) · d(-1) and 1.0 g · kg(-1) · d(-1) of C. frondosa powder suspensions. The degrees of hepatocyte fatty degeneration and inflammation were observed under the optical microscope with routine HE staining. The NAFLD activity scores (NAS) were calculated. Serum ALT, AST and hepatic TG and CHOL were tested by the biochemical method. The hepatic MDA was examined by thiobarbituric acid method. The hepatic SOD was tested by the xanthine oxidase test. The hepatic GSH-PX activity was determined by the dithio-nitrobenzoic acid method. Hepatic TNF-α and IL-6 were detected by the enzyme-linked immunosorbent assay (ELISA). The NASH model group induced by high-fat diets showed higher hepatic NAS, ser- um ALT, AST, CHOL and hepatic TG, CHOL, MDA, TNF-α, IL-6 (P < 0.01 or P < 0.05) and lower serum TG and hepatic SOD, GSH-PX (P < 0.01, P < 0.05) than the normal control group. After being intervened with different doses of G. frondosa, the NASH group revealed significantly lower hepatic NAS, serum ALT and hepatic TG, CHOL, MDA, TNF-α and IL-6 (P < 0.05) and higher hepatic SOD, GSH-PX (P < 0.05) than the model group. G. frondosa may prevent the further development of NASH by improving the disorder of lipid metabolism in rats with NASH induced by high-fat diets, relieving the level of oxidative stress and reducing the generation of inflammatory cytokines.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Grifola ; chemistry ; Humans ; Interleukin-6 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

Objective To observe the effect of smoking on the great saphenous vein tension. Methods The rings of great saphenous vein in 3mm long of selected from 31 patients with coronary artery bypass grafting were divided into three groups :smoking over 10 years( group A, n = 12 ), ex-smoking over 1 years ( group B, n =9 ) and non-smoking( group C, n =10). The changes of the tone were measured in organ chamber at 37℃ with a constant supply of oxygen when vasoconstriction induced by phenylephrine ( 10 -9 - 10 -5 mmol/L), and vasodilatation by acetylcholine ( 10 -9 - 10 -5 mmol/L) or nitroglycerin( 10 -9 - 10 -4 mmol/L)after the rings were precontracted by 10 -5 mmol/L phenylephrine. Results Vasoconstriction induced by phenylephrine and vasodilatation by nitroglycerin is no significant difference among three groups. Compared with group A, vasodilatation by acetylcholine was significantly increased in group B or C, while there is no significant difference between group B and C. Conclusion Smoking has a deleterious effect on the endothelial function of great saphenous vein, however, smoking cessation over 1 year may help to restore the endothelial function impaired by smoking.

Objective To explore the risk factors of hyperbilirubinemia in late preterm infants. Methods Clinical data of 211 cases of late preterm infants with hyperbilirubinemia and 246 cases of late preterm infants without hyperbilirubinemia were retro-spectively analyzed between 2011 and 2012. The risk factors of hyperbilirubinemia were filtered. Results Twenty-seven cases of late premature infants with hyperbilirubinemia were severe. Hospital stay less than 3 days, birth asphyxia history, small for gestatio-nal age, head hematoma, delivery injury, hypoalbuminemia, polycythemia, infection, hemolytic disease, feeding intolerance, and fe-tal excretion delay were associated with hyperbilirubinemia (P<0.05). Rural origin, pregnancy-induced hypertension syndrome and premature rupture of membrane were also associated with hyperbilirubinemia (P<0.05). Multivariate logistic regression analysis showed the history of birth asphyxia , fetal excretion delay, hypoalbuminemia, pregnancy-induced hypertension syndrome were risk factors of hyperbilirubinemia in late preterm infants (OR=2.35-4.05). Pregnancy-induced hypertension syndrome and hemolytic dis-ease were risk factors of severe hyperbilirubinemia in late preterm infants (OR=5.74, 73.64). Conclusions Neonatal asphyxia, fetal excretion delay, hypoalbuminemia and pregnancy-induced hypertension syndrome are risk factors of hyperbilirubinemia in late pre-term infants. Strengthening the management of pregnancy-induced hypertension syndrome and the treatment of newborn hemolytic disease can reduce the occurrence of severe hyperbilirubinemia in late preterm infants.

OBJECTIVE To study how to properly evaluate the curative effect of chronic sinusitis and nasal polyps management with endoscopic sinus surgery(ESS). METHODS Nasal airway resistance, olfactory function, the morphological character of mucosa in nasal and sinus cavity after ESS were surveyed by anterior rhinomanometry,T&T olfactometer standard odors for measuring olfactory sense,acoustic rhinometry and scoring measure of mucosa. RESULTS After ESS, nasal airway resistance decreased and olfactory functions improved obviously. The morphological characters of mucosa in nasal and sinus cavity affect the surgical result directly. CONCLUSION As the determining methods of nasal function after/before ESS,anterior rhinomanometry,T&T olfactometer standard odors,acoustic rhinometry and scoring measure of mucosa can be used to comprehensive estimate the curative effect of ESS objectively.